杏褪绿卷叶植原体新疆分离物核糖体蛋白基因的克隆与序列分析  

Cloning and sequencing of the ribosomal protein gene of the phytoplasma of apricot chlorotic leaf roll in Xinjiang

作　　者：韩剑[1,2] 陈杭 艾克热木·买买提 罗明 唐章虎[3] HAN Jian;CHEN Hang;AIKEREMU·Maimaiti;LUO Ming;TANG Zhanghu(Agronomy Faculty,Xinjiang Agricultural University,Urumqi,Xinjiang 830052,China;Key Laboratory of Harmful Biological Monitoring and Safety Prevention of Xinjiang Autonomous Region Forestry Universities,Urumqi,Xinjiang 830052,China;Luntai National Pomology Germplasm Garden,Academy of Agricultural Science of Xinjiang,Luntai,Xinjiang 841600,China)

机构地区：[1]新疆农业大学农学院,新疆乌鲁木齐830052 [2]新疆自治区高校农林有害生物监测与安全防控重点实验室,新疆乌鲁木齐830052 [3]新疆农业科学院轮台国家果树资源圃,新疆轮台841600

出　　处：《生物安全学报》2021年第1期36-42,共7页Journal of biosafety

基　　金：国家自然科学基金(31600011);新疆维吾尔自治区研究生教育创新计划科研创新项目(XJGRI2017-072)。

摘　　要：【目的】了解杏褪绿卷叶植原体新疆分离物的系统发育关系及遗传分化,确定其分类地位。【方法】利用植原体核糖体蛋白(rp)基因的特异性引物rpF1/rpR1对新疆轮台县托克逊县杏褪绿卷叶病植株总DNA进行PCR扩增,并对部分扩增片段克隆、测序及序列分析。【结果】获得杏褪绿卷叶植原体新疆分离物rp基因片段大小为1196 bp,该片段包含部分rpS19以及rpL22和rpS3基因的全部序列。序列相似性和系统进化分析表明,杏褪绿卷叶植原体新疆分离物与16SrⅤ-rp亚组中的各代表性植原体的rp基因核苷酸序列相似性达到95.7%~99.3%,其中与rpⅤ-C亚组的甜樱桃绿化植原体和枣疯病植原体的相似性最高,核苷酸及氨基酸相似性分别达到99.2%~99.3%和98.3%~98.4%。进一步虚拟RFLP分析,发现杏褪绿卷叶植原新疆分离物rp基因的酶切图谱与rpⅤ-C亚组成员相似性最高,但在MseⅠ、SspⅠ和TaqⅠ的酶切位点上存在差异。综上初步判断其可能属于16SrⅤ组(榆树黄化组)中的一个新rp亚组。【结论】本研究首次报道了杏褪绿卷叶植原体新疆分离物的rp基因序列,确定了其分类地位,为杏褪绿卷叶病的早期诊断和检测提供了基础。【Aim】To examine the molecular phylogenetic relationship,genetic differentiation and taxonomic status of apricot chlorotic leaf roll(ACLR)phytoplasmas from Xinjiang isolates.【Method】The ribosomal protein gene(rp)of the phytoplasma of apricot chlorotic leaf roll was amplified by PCR with the specific primer pairs rpF1/rpR1 in the diseased apricot sample from Luntai County and Tuokexun County in Xinjiang Autonomous Regions,then the partial rp gene was cloned,sequenced and analyzed with other phytoplasmas.【Result】The results showed that a 1196 bp DNA fragment was amplified from the infected apricot.The fragment included partial rpS19 gene,complete rpL22 and rpS3 genes.The similarity and phylogenetic analyses showed that the sequence shared 95.7%-99.3% similarity with those of representative phytoplasmas in the 16SrⅤ-rp subgroup.The NCBI blast results showed that the genetic distance of ACLR-Xinjiang isolates was closely related to the rpV-C subgroup,which shared the highest homology of nucleotide and amino acid sequences of 99.2%-99.3% and 98.3%-98.4% with the strains of prunus avium virescence phytoplasma and jujube witches′-broom phytoplasma.Furthermore,virtual RFLP analysis showed that the phytoplasma might belonged to a new subgroup within the elm yellows group(16SrV).The most similar was the reference pattern of the rpV-C subgroup(AB052876),but showed significant differences in the profile with MseⅠ,SspⅠand TaqⅠ.【Conclusion】This study reported the rp gene sequences of ACLR phytoplasmas and classified phytoplasma of ACLR in Xinjiang for the first time.It provides the basis for early diagnosis and detection.

关 键 词：杏 杏褪绿卷叶病 植原体 核糖体蛋白基因 序列分析 

分 类 号：S436.629[农业科学—农业昆虫与害虫防治]